Antifreeze proteins from the sea raven, Hemitripterus americanus. Further evidence for diversity among fish polypeptide antifreezes.
The antifreeze proteins of the sea raven, Hemitripterus americanus, were isolated and compared with other fish antifreeze proteins. The sea raven contains one major protein of molecular weight 14,000-16,000 with little or no carbohydrate. Except for its similar seasonal appearance, the sea raven antifreeze protein differs from other polypeptide antifreeze in its amino acid composition, secondary structure, and immunological specificity. Amino acid analysis of sea raven antifreeze showed that it contains a high amount of half-cystine, hydrophilic amino acids, and only an average amount of alanine. In contrast, all other fish antifreeze proteins contain approximately 60% alanine and no half-cystine residues. Furthermore, the sea raven antifreeze protein is sensitive to sulfhydryl reagents. The antifreeze activity was decreased by 67% in the presence of 0.01 M dithiothreitol. Circular dichroism studies indicated the absence of significant amounts of alpha-helix and the possible presence of beta-structure. Antibodies raised against the antifreeze protein did not cross-react with the known polypeptide antifreeze from the winter flounder and shorthorn sculpin (Hew, C. L., Fletcher, G. L., and Ananthanarayanan, V. S. (1980) Can. J. Biochem. 58, 377-383). A specific radioimmunoassay was developed for the sea raven antifreeze protein and was used to quantitate the protein concentration in the fish. The seasonal profile obtained by radioimmunoassay was compatible with the antifreeze activity determined with a freezing point osmometer.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002942:Circular Dichroism; D005399:Fishes / Q000097:blood*; D005779:Immunodiffusion; D007700:Kinetics; D010455:Peptides / Q000097:blood*; D011487:Protein Conformation; D011489:Protein Denaturation; D011863:Radioimmunoassay; D013045:Species Specificity; D013696:Temperature
D000596:Amino Acids; D010455:Peptides